CN102876603B - Bacillus amyloliquefaciens Mg116 and application thereof - Google Patents

Bacillus amyloliquefaciens Mg116 and application thereof Download PDF

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CN102876603B
CN102876603B CN2012103546282A CN201210354628A CN102876603B CN 102876603 B CN102876603 B CN 102876603B CN 2012103546282 A CN2012103546282 A CN 2012103546282A CN 201210354628 A CN201210354628 A CN 201210354628A CN 102876603 B CN102876603 B CN 102876603B
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bacillus amyloliquefaciens
bacterial strain
suspension agent
suspension
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CN102876603A (en
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庄义庆
杨敬辉
吴琴燕
陈宏州
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JIANGXI NEW DRAGON BIOTECHNOLOGY Co.,Ltd.
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Zhenjiang Institute of Agricultural Sciences Jiangsu Hilly Area
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Abstract

The invention relates to bacillus amyloliquefaciens Mg116 and a method for preparing a suspending agent from bacillus amyloliquefaciens Mg116 fermentation broth. The bacillus amyloliquefaciens Mg116 has the collection number of No.3967 in the CGMCC (China General Microbiological Culture Collection Center). The preparation can be used for effectively preventing and treating the grape, watermelon and strawberry anthracnose caused by glomerella cingulata which is a plant pathogenic fungus.

Description

A kind of bacillus amyloliquefaciens Mg116 bacterial strain and purposes
Invention field
The present invention relates to a kind of microorganism and purposes, the preparation method more specifically to a kind of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Mg116 bacterial strain, purposes and preparation thereof, belong to biological pesticide technical field.
Background technology
Enclosing small cluster shell bacterium (Glomerella cingulata) by plant pathogenic fungi, to infect the anthrax of initiation be Major Diseases on the cash crop such as strawberry, grape and watermelon,, to the height of anthrax prevention effect, the income at crop yield and plantation family will be had influence on directly.The control of anthrax is current mainly carrys out prevention and control by chemical prevention.Mainly use following Agro-chemicals control: Azoxystrobin (azoxystrobin), kresoxim-methyl (kresoxim-methyl), difenoconazole (difenoconazole), enostroburin (enestroburin), boscalid amine (boscalid), derosal (carbendazim), pyraclostrobin (pyraclostrobin) and prochloraz (prochloraz) etc.For a long time,, because multifrequency is inferior, multiple doses is used chemical agent, cause anthrax bacteria all to produce anti-(anti-) property of medicine in various degree to chemical agent commonly used, occur repeatedly the example that control is failed in production.
The biological pesticide special preparation that successfully is used at present the control anthrax bacteria does not also have.Document has reported that the biological and ecological methods to prevent plant disease, pests, and erosion Pseudomonas (kind) of biological control control disease mainly contains viride (Trichoderma viride) and the trichoderma harziarum (Trichoderma harzianum) in (1) trichoderma fungi (Trichoderma spp.); (2) subtilis (Bacillus subtilis) in bacillus (Bacillus spp.), bacillus polymyxa (Bacillus polymyxa) and bacillus megaterium (Bacillus megaterium); (3) the false single-cell bacteria of the fluorescence in Rhodopseudomonas bacterium (Pseudomonas spp.) (Pseudomonas fluorescens) etc.
Although the host range of anthrax bacteria is very wide, important host crop is garden crop, as strawberry, grape and watermelon etc.Such crop is mostly the fruits in season class, and be mostly namely adopt instant, so the selection of medicament is had significant limitation.Therefore the biological control of anthrax had certain active demand on producing.The bacterium that flys up and down that can effectively prevent and treat anthrax provided by the invention, do not mention in the prior art and disclose.
Summary of the invention
The present invention solves the deficiency that prior art exists, provide a strain the new genus bacillus that separates: bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Mg116 bacterial strain, this bacterial strain can be used for control and infect the anthrax of initiation by enclosing small cluster shell bacterium (Glomerella cingulata).The present invention simultaneously also provides the preparation method who is processed into suspension agent with this bacillus amyloliquefaciens Mg116 bacterial strain.
The present invention is achieved by the following technical solutions:
Bacillus amyloliquefaciens of the present invention (Latin name is Bacillus amyloliquefaciens) Mg116 bacterial strain, on July 8th, 2010 in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, the depositary institution address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, and preserving number is: CGMCC No. 3967.Bacterial classification of the present invention has following feature:
(1) cellular form of Mg116 bacterial strain and physiological and biochemical property (in Table 1)
Table 1, Mg116 strain cell form and physicochemical characteristics
Figure GDA0000225984591
(2) the 16S rRNA gene order of bacillus amyloliquefaciens Mg116 bacterial strain of the present invention is:
CTCCATAAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCAGCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGAACAGATTTGTGGGATTGGCTTAACCTCGCGGTTTCGCTGCCCTTTGTTCTGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCACCTTAGAGTGCCCAACTGAATGCTGGCAACTAAGATCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACTCTGCCCCCGAAGGGGACGTCCTATCTCTAGGATTGTCAGAGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTAAGGGGCGGAAACCCCCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGAGCCCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTGCCGCCCTATTTGAACGGCACTTGTTCTTCCCTAACAACAGAGCTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCCGAAGCCACCTTTTATGTCTGAACCATGCGGTTCAGACAACCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACATCAGGGAGCAAGCTCCCATCTGTCCGCTCGA。
With the biocontrol fungicide of preventing and treating the bacterial strain Mg116 preparation of anthrax of the present invention, its preparation method comprises the following steps:
(A) acquisition of bacterium liquid
With aseptic bud label picking streak culture good Mg116 bacterial strain list bacterium colony spot on the PDA solid medium in advance, being inoculated in the volume that 5 mL PDA liquid mediums are housed is in the triangular flask of 20 mL, shaking culture 16 h under 30 ℃, 200 rpm conditions, spend the night, it is in the triangular flask of 1000 mL that gained nutrient solution 5 mL of upper step all are inoculated in the volume that 400 mL PDA nutrient solutions are housed, shaking culture 16 h under 200 rpm, 30 ℃ of conditions; The 400mL nutrient solution of gained is inoculated in the fermentor tank that the volume that 20 L nutrient solutions are housed is 30L (east, Zhenjiang GUS-30), and in the 20L fermentation culture, solid tolerant content is: dregs of beans 100 g, yam starch 200 g, sucrose 25 g, yeast powder 25 g, CaCO 320 g, MnSO 41.0 g arranges fermentation condition and is: dissolved oxygen 100%, stirring velocity are 350 rpm, 30 ℃ of leavening temperatures, fermentation time 36 h, pH 7.0-7.2;
(B) microbial inoculum complete processing
The fermented liquid that ferments is processed into suspension agent as follows:
(1) fermented liquid centrifugal (8000 rpm) is rear with 20% glycerol solution (V/V) (solution is prepared with sterilized water) Eddy diffusion, measure the living spores content of suspension, regulate the living spores number of suspension with 20% glycerol solution (V/V) according to measurement result, it is 1.0 * 10 that whole content is adjusted to gemma content 10Cfu/mL(is every milliliter and contains 10,000,000,000 living spores); (2) in the above-mentioned suspension that regulates by 5 g/L(W/V) consumption add xanthan gum, and stir, the concentration that obtains the finished product suspension agent is 10,000,000,000 living spores/mL.
The invention has the beneficial effects as follows:
(1) the invention provides the new Biocontrol Bacillus of a strain---bacillus amyloliquefaciens MG116 bacterial strain, the suspension agent that is made by this strain fermentation, can be used for control and infect the anthracnose of crop of initiation by enclosing small cluster shell bacterium (Glomerella cingulata).Field demonstration is prevented and treated result and is shown, this microbial inoculum all has good prevention effect to Plant diseasess such as Strawberry anthracnose, bitter rot or anthracnose of grape and watermelon anthrax diseases;
(2) the microbial inoculum storing property is stable, and prevention effect is good, and the zymotechnique of scale operation is simple, low production cost.
Embodiment
Embodiment
Bacillus amyloliquefaciens of the present invention (Latin name is Bacillus amyloliquefaciens) bacterial strain Mg116, on July 8th, 2010 in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, preserving number is: CGMCC No. 3967.The cellular form of bacterial strain of the present invention and physiological and biochemical property are in Table 1.
The 16S rRNA gene order of the bacillus amyloliquefaciens Mg116 bacterial strain of the present embodiment is:
CTCCATAAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCAGCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGAACAGATTTGTGGGATTGGCTTAACCTCGCGGTTTCGCTGCCCTTTGTTCTGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCACCTTAGAGTGCCCAACTGAATGCTGGCAACTAAGATCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACTCTGCCCCCGAAGGGGACGTCCTATCTCTAGGATTGTCAGAGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTAAGGGGCGGAAACCCCCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGAGCCCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTGCCGCCCTATTTGAACGGCACTTGTTCTTCCCTAACAACAGAGCTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCCGAAGCCACCTTTTATGTCTGAACCATGCGGTTCAGACAACCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACATCAGGGAGCAAGCTCCCATCTGTCCGCTCGA。
With the biocontrol fungicide of preventing and treating the Mg116 bacterial strain preparation of anthrax of the present invention, its preparation method comprises the following steps:
(A) acquisition of bacterium liquid
With aseptic bud label picking streak culture good Mg116 bacterial strain list bacterium colony spot on the PDA solid medium in advance, being inoculated in the volume that 5 mL PDA liquid mediums are housed is in the triangular flask of 20 mL, shaking culture 16 h under 30 ℃, 200 rpm conditions, spend the night, it is in the triangular flask of 1000 mL that gained nutrient solution 5 mL of upper step all are inoculated in the volume that 400 mL PDA nutrient solutions are housed, shaking culture 16 h under 200 rpm, 30 ℃ of conditions; The 400mL nutrient solution of gained is inoculated in the fermentor tank that the volume that 20 L nutrient solutions are housed is 30L (east, Zhenjiang GUS-30), and in the 20L fermentation culture, solid tolerant content is: dregs of beans 100 g, yam starch 200 g, sucrose 25 g, yeast powder 25 g, CaCO 320 g, MnSO 41.0 g arranges fermentation condition and is: dissolved oxygen 100%, stirring velocity are 350 rpm, 30 ℃ of leavening temperatures, fermentation time 36 h, pH 7.0-7.2;
(B) microbial inoculum complete processing
The fermented liquid that ferments is processed into suspension agent as follows:
(1) fermented liquid centrifugal (8000 rpm) is rear with 20% glycerol solution (V/V) (solution is prepared with sterilized water) Eddy diffusion, measure the living spores content of suspension, regulate the living spores number of suspension with 20% glycerol solution (V/V) according to measurement result, it is 1.0 * 10 that whole content is adjusted to gemma content 10Cfu/mL(is every milliliter and contains 10,000,000,000 living spores); (2) in the above-mentioned suspension that regulates by 5 g/L(W/V) consumption add xanthan gum, and stir, the concentration that obtains the finished product suspension agent is 10,000,000,000 living spores/mL.
Implementation result
(1) Mg116 bacterial strain fermentation liquor processing and suspension agent to enclosing the indoor virulence of small cluster shell bacterium (Glomerella cingulata)
Mg116 bacterial strain fermentation liquor processing and suspension agent as follows to the Toxicity Determination method of enclosing the small cluster shell bacterium:
With Mg116 bacterial strain suspension agent (10,000,000,000 living spores/mL) be mixed with and contain 10 8, 10 7, 10 6, 10 5, 10 4The PDA of cfu/mL is dull and stereotyped, and to establish sterilized water be blank; (10,000,000,000 living spores/mL) measure by above-mentioned same concentrations by wettable powder with subtilis for the contrast medicament.Be the punch tool of 4 mm with diameter, beat and get the bacterium cake on the concentric(al) circles at the edge that encloses small cluster shell bacterium flat board of having grown in advance, to containing the above-mentioned central authorities that contain the bacterium flat board, each concentration of each bacterial strain is established 4 repetitions with pure culture biscuits involvng inoculation.Flat board is placed in 25 ℃ of heat insulating culture casees, cultivates 96 h.Measure the clean growth diameter of bacterium colony (the clean growth diameter=colony diameter of bacterium colony-bacterium cake diameter) with the right-angled intersection method, calculate inhibiting rate.Inhibiting rate=(the clean growth diameter of blank bacterium colony-clean growth diameter of pastille the bacterium colony)/clean growth diameter of blank bacterium colony * 100%.With DPS 13.0 computed in software virulence regression equations.
The Mg116 suspension agent is to enclosing the Toxicity Determination result of small cluster shell bacterium:
The toxicity test result shows, the Mg116 suspension agent all has strongly inhibited effect (table 2,3,4), its EC to the growth of enclosing small cluster shell bacterium mycelia 50Value is 7.6048 * 10 3Cfu/mL.By contrast, under identical extension rate condition the subtilis wettable powder to enclosing the EC of small cluster shell bacterium 50Value is respectively 1.8651 * 10 6Cfu/mL.The antibacterial virulence utmost point of Mg116 suspension agent is significantly higher than the subtilis wettable powder.
Table 2. Mg116 suspension agent is to enclosing the average inhibiting rate of small cluster shell bacterium
Dilution gradient (cfu/mL) 10 8 10 7 10 6 10 5 10 4
Average inhibiting rate (%) 100 89.55 75.47 66.12 53.85
Table 3. subtilis wettable powder is to enclosing the average inhibiting rate of small cluster shell bacterium
Dilution gradient (cfu/mL) 10 8 10 7 10 6 10 5 10 4
Average inhibiting rate (%) 70.75 56.45 42.71 30.54 19.55
Table 4. Mg116 suspension agent is to enclosing the toxicity test result of small cluster shell bacterium
(2) field controling test of Mg116 suspension agent to Strawberry anthracnose
The test treatment process: this test arrangement carries out in the strawberry of the Houbai Town, Jurong City, Jiangsu Province agriculture factory field of growing seedlings, and strawberry cultivars is red cheek (high sense anthrax).The control of spraying for the first time on June 10th, 2011, interval spray in 10 days once, control 8 times continuously altogether, September 1 the investigation prevention effect.Test medicine is processed: (1) bacillus amyloliquefaciens Mg116 suspension agent (1000 times of liquid of 10,000,000,000 living spores/mL); (2) the contrast medicament is with subtilis wettable powder (1000 times of liquid of 10,000,000,000 living spores/mL); (3) blank is sprayed with clear water.Mu water consumption 50 kg, the residential quarter area is 10 m 2.Test is established 4 and is repeated.
Result: field test results (table 5) shows, the prevention effect of 1000 times of liquid of Mg116 suspension agent is 95.60%, and the prevention effect of 1000 times of liquid of subtilis wettable powder is only to reach the statistics level difference between 81.33%, two medicament by contrast.
The field controling test result of table 5 Mg116 suspension agent to Strawberry anthracnose
Annotate: different capitalizations represent P＜0.01.
(3) field controling test of Mg116 suspension agent to bitter rot or anthracnose of grape
The test treatment process: be popular in agricultural science and technology Demonstration Garden (Houbai Town, Jurong City, Jiangsu Province) in the Wanshan Mountain, Jiangsu Province, test garden grape contiguous plant, grape variety is huge rich.Adopt the horizontal rack formula to plant, distance between rows and hills 2 m * 4 m, age of tree life in 6 years.Horse liver soil, middle fertility, organic content 1% left and right, orchard management more solito.Duration of test (6～September in 2011).Test medicine is treated to: 1000 times of liquid of (1) Mg116 suspension agent; (2) the contrast medicament is 1000 times of liquid of subtilis wettable powder; (3) blank is not for preventing and treating.Residential quarter area 800 m 2, random district group is arranged, 4 repetitions.Test in the 1st spray medicine on June 20, prevent and treat altogether 5 times every 10 days (adjusting the spray medicine date while running into rainy day) later, last 1 control date is August 13.Adopt Backpack electrostatic atomizer even spraying, every 667m 2With amount of liquid medicine 100 kg.Investigation on 14 days (August 27) preventive effect after last 1 dispenser.Select at random 4 strain grapes during investigation in the middle of each residential quarter, in upper, middle and lower, left and right 5 orientation get respectively 1 fruit ear, totally 20 fruit ears, put down in writing the sickness rate of various diseases in each fruit ear axle, fruit grain and carpopodium, calculates that disease refers to and prevention effect.
Result: field test results (table 6) shows, the prevention effect of 1000 times of liquid of Mg116 suspension agent is 90.72%, and the prevention effect of 1000 times of liquid of subtilis wettable powder is only to reach the statistics level difference between 48.44%, two medicament by contrast.
The field controling test result of table 6 Mg116 suspension agent to bitter rot or anthracnose of grape
Figure GDA0000225984594
Annotate: different capitalizations represent P＜0.01.
(4) field controling test of Mg116 suspension agent to watermelon anthrax
The test treatment process: this test arrangement carries out in the greenhouse watermelon field of Houbai Town, Jurong City, Jiangsu Province agriculture factory, and variety of watermelon is No. 3, Dongting Lake.The control of spraying for the first time on May 20th, 2011, interval spray in 10 days once, control 4 times continuously altogether, June 30 the investigation prevention effect.Investigational agent is processed: (1) bacillus amyloliquefaciens Mg116 suspension agent (1000 times of liquid of 10,000,000,000 living spores/mL); (2) the contrast medicament is with subtilis wettable powder (1000 times of liquid of 10,000,000,000 living spores/mL); (3) blank is sprayed with clear water.Mu water consumption 50 kg, the residential quarter area is 50 m 2.Test is established 4 and is repeated.
Result: field test results (table 7) shows, the prevention effect of 1000 times of liquid of Mg116 suspension agent is 93.5%, and the prevention effect of 1000 times of liquid of subtilis wettable powder is only to reach the statistics level difference between 44.46%, two medicament by contrast;
The field controling test result of table 7 Mg116 suspension agent to bitter rot or anthracnose of grape
Figure GDA0000225984595
Annotate: different capitalizations represent P＜0.01.

Claims (3)

  1. A bacillus amyloliquefaciens ( Bacillus amyloliquefaciens) the Mg116 bacterial strain, this bacterial classification is in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, and preserving number is: CGMCC No. 3967.
  2. 2. bacillus amyloliquefaciens Mg116 bacterial strain according to claim 1 is characterized in that the 16S rRNA gene order of this bacterial strain is:
    CTCCATAAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCAGCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGAACAGATTTGTGGGATTGGCTTAACCTCGCGGTTTCGCTGCCCTTTGTTCTGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCACCTTAGAGTGCCCAACTGAATGCTGGCAACTAAGATCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACTCTGCCCCCGAAGGGGACGTCCTATCTCTAGGATTGTCAGAGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTAAGGGGCGGAAACCCCCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGAGCCCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTGCCGCCCTATTTGAACGGCACTTGTTCTTCCCTAACAACAGAGCTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCCGAAGCCACCTTTTATGTCTGAACCATGCGGTTCAGACAACCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACATCAGGGAGCAAGCTCCCATCTGTCCGCTCGA。
  3. 3. the application of bacillus amyloliquefaciens Mg116 bacterial strain claimed in claim 1 aspect control Strawberry anthracnose, bitter rot or anthracnose of grape and watermelon anthrax.
    4. suspension agent that is prepared from the fermented liquid that the described bacillus amyloliquefaciens Mg116 of claim 1 strain fermentation obtains.
    5. method for preparing suspension agent claimed in claim 4 is characterized in that comprising the following steps:
    (A) acquisition of bacterium liquid
    With aseptic toothpick picking streak culture good Mg116 bacterial strain list bacterium colony spot on the PDA solid medium in advance, being inoculated in the volume that 5 mL PDA liquid mediums are housed is in the triangular flask of 20 mL, shaking culture 16 h under 30 ℃, 200 rpm conditions, it is in the triangular flask of 1000 mL that gained nutrient solution 5 mL of upper step all are inoculated in the volume that 400 mL PDA nutrient solutions are housed, shaking culture 16 h under 200 rpm, 30 ℃ of conditions; Again 400 mL nutrient solutions of gained being inoculated in the volume that 20 L nutrient solutions are housed is in the fermentor tank of 30L, and in 20 L fermentation cultures, solid tolerant content is: dregs of beans 100 g, yam starch 200 g, sucrose 25 g, yeast powder 25 g, calcium carbonate 20 g, MnSO 41.0 g arranges fermentation condition and is: dissolved oxygen 100%, stirring velocity are 350 rpm, 30 ℃ of leavening temperatures, fermentation time 36 h, pH 7.0-7.2;
    (B) microbial inoculum complete processing
    The fermented liquid that ferments is processed into suspension agent as follows:
    (1) fermented liquid is being removed supernatant after centrifugal 10 min under 8000 rpm conditions, the solution Eddy diffusion that includes 20% glycerol that excess is prepared with sterilized water, measure the living spores content of suspension, regulate the living spores number of suspension with 20% glycerol solution according to measurement result, it is 1.0 * 10 that whole content is adjusted to gemma content 10Cfu/mL, the unit ratio of above-mentioned glycerol is V/V; (2) consumption by 5 g/L adds xanthan gum in the above-mentioned suspension that regulates, and stirs, and the concentration that obtains the finished product suspension agent is 1.0 * 10 10Cfu/mL, wherein the unit ratio of xanthan gum content is W/V.
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